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低温顕微鏡と高速ビデオカメラを組み合わせた細胞内凍結過程の解析 Analysis of Intracellular Freezing Process of Plant Cells with Cryomicroscope and High-Speed Video Camera 上村, 松生 中川原, 千早 河村, 幸男 吉田, 静夫 江藤, 剛治 竹原, 幸生 低温生物工学会 ご利用に当たっては「著作権法」に従うことをお願いいたします． Crymicroscopy Intracellular freezing High-speed video camera Protoplast Plasma membrane 低温顕微鏡 細胞内凍結 高速ビデオカメラ プロトプラスト 細胞膜 Normal-speed (32 frames/sec) and high-speed (up to 4,000 frames/sec) video cameras attached with a cryomicroscope were utilized to analyze the freezing process of protoplasts isolated from Arabidopsis leaves and Jerusalem artichoke tubers. With protoplasts isolated from leaves of non-acclimated Arabidopsis seedlings, intracellular freezing was observed even when protoplasts were frozen to -10℃ at rates of 0.5 to 1.0℃/min, and the frequency increased significantly at 2.5 to 10℃/min. After cold acclimation for 1 day at 2℃, the intracellular freezing did not occur at cooling rates of 1.0℃ or slower. Cold acclimation for 2 to 7 days further resulted in a shift of the temperature range over which intracellular freezing occurred to lower temperatures. High-speed video microscopy revealed that intracellular freezing was initiated, with no exceptions, at places near the plasma membrane. However, it was not possible with techniques used in the present study to determine whether ice formation initially occurs inside or outside the cell. Nevertheless, these studies demonstrate that videomicroscopy combined with cryostage provides an insight to understand the effect of cold acclimation on the freezing process of plant protoplasts. 低温生物工学会 2002-12-28 jpn journal article VoR https://iwate-u.repo.nii.ac.jp/records/9240 1340-7902 低温生物工学会誌 48 2 129 133 https://iwate-u.repo.nii.ac.jp/record/9240/files/cc-v48n2p129-133.pdf application/pdf 441.2 kB 2016-11-14